Isolation of a soil bacterium capable of biodegradation and detoxification of endosulfan and endosulfan sulfate

Endosulfan, an endocrine disrupting chemical, is a widely used cyclodiene organochlorine pesticide worldwide, and it blocks neuronal GABA(A)-gated chloride channels in mammals and aquatic organisms. Endosulfan and its metabolites, such as endosulfan sulfate, are persistent in environments and are considered as toxic chemicals. For bioremediation of endosulfan, in this study, an attempt was made to isolate an endosulfan and endosulfan sulfate degrading bacterium from endosulfan-polluted agricultural soil. Through repetitive enrichment and successive subculture using endosulfan or endosulfan sulfate as the sole carbon source, a bacterium KS-2P was isolated. The KS-2P was identified as Pseudomonas sp. on the basis of the results of a 16S rDNA sequencing analysis and MIDI test. The degradation ratios for endosulfan or endosulfan sulfate in minimal medium containing endosulfan (23.5 microg mL(-1)) or endosulfan sulfate (21 microg mL(-1)) were 52% and 71%, respectively. Our results suggest that Pseudomonas sp. KS-2P has potential as a biocatalyst for endosulfan bioremediation.     

Comparative study of methods for the extraction of eleven organophosphorus pesticide residues in rice.

Several extraction methods are compared for the simultaneous analysis of organophosphorus pesticides in unpolished rice. Four stationary phases were used for the subsequent gas-liquid chromatographic (GLC) determination of the selected pesticides. Using 3 different GLC columns, 11 pesticides were completely separated and identified. The efficiency of the cleanup and the sensitivity of the analytical method were evaluated by using powdered unpolished rice samples fortified with the pesticides and also wheat and dried bean samples. Average recoveries ranged from 74.7% for disulfoton to 97.4% for malathion in unpolished rice and from 68.1% for disulfoton to 108.3% for malathion in other crops. The method described is applicable to the analysis of selected organophosphorus pesticide residues in unpolished rice, wheat, buckwheat, and dried beans.     

Determination of organochlorine and organophosphorus pesticide residues in eggs using a solid phase extraction cleanup

A multiresidue solid phase extraction (SPE) method for the isolation and subsequent gas chromatographic determination of nonpolar organochlorine and polar organophosphorus pesticide residues in eggs is described. The method uses an acetonitrile extraction followed by an SPE cleanup using graphitized carbon black and aminopropyl SPE columns. Organophosphorus pesticides are determined by gas chromatography with flame photometric detection. After further cleanup of the extract using Florisil SPE columns, organochlorine pesticides are determined by gas chromatography with electron capture detection. Studies were performed using eggs containing both fortified and incurred pesticide residues. The average recoveries were 86-108% for 8 fortified organochlorine pesticide residues and 61-149% for 28 fortified organophosphorus pesticide residues.     

Development of green onion and cabbage certified reference materials for quantification of organophosphorus and pyrethroid pesticides

Green onion and cabbage certified reference materials for the analysis of pesticide residues were issued by the National Metrology Institute of Japan, part of the National Institute of Advanced Industrial Science and Technology. Green onion and cabbage samples were grown so as to contain several kinds of organophosphorus and pyrethroid pesticides, and those were collected from a field in the Kochi Prefecture in Japan. The certification was carried out by using multiple analytical methods to ensure the reliability of analytical results; the values of target pesticides (diazinon, fenitrothion, cypermethrin, etofenprox, and permethrin for green onion and chlorpyrifos, fenitrothion, and permethrin for cabbage) were obtained by isotope dilution mass spectrometry. Certified values of target pesticides were 0.96-13.9 and 2.41-6.9 mg/kg for green onion and cabbage, respectively. These are the first green onion and cabbage powder certified reference materials in which organophosphorus and pyrethroid pesticides are determined.     

采用MECC在线堆积同步检测茶叶中七种残留农药

为解决茶叶中的痕量农药残留的快速检测问题,建立了一种基于胶束毛细管电泳在线堆积技术的茶叶中7种拟除虫菊酯类农药痕量残留的一次进样同步检测方法.通过参数优化,得到最优的柱上在线堆积浓缩操作工艺:以15%异丙醇、20%乙腈、60 mmol/L十二烷基硫酸钠(SDS)和50 mmol/L tris的混合物为背景缓冲溶液(BGS).茶叶的浸提液经过固相萃取净化吹干后,用1 mmol.L SDS溶解溶液残渣.进样100 s后施加反向电压,进行农药反向在线堆积,当电流恢复至正常分离时的95%时改变电压方向,进行正常分离.经过在线堆积浓缩,7种拟除虫菊酯类农药痕量残留堆积因子均高于17,回收率在82%以上,检测下限分别达到:功夫菊酯0.05 mg/kg、联苯菊酯0.05 mg/kg、百树菊酯0.05 mg/kg、高效氯氰菊酯0.05 mg/kg、溴氰菊酯0.2 mg/kg、二氯菊酯0.05 mg/kg、氰戊菊酯0.05 mg/kg,满足了2006年欧盟茶叶中拟除虫菊酯类农药残留的最低标准.     

巢湖东半湖沉积物中有机氯农药的残留特征及风险评价

采样测定了巢湖东半湖4个样点的表层沉积物中有机氯农药（OCPs）的含量。结果表明,11种有机氯农药在样品中被检出,总含量为8.26～31.73 ng.g^-1;OCPs在沉积物中的垂直分布从上往下大体呈递减趋势;且OCPs的最高含量都出现在上层沉积物中,说明巢湖东半湖沉积物中有机氯农药主要集中在0-3 cm的表层。根据分析,DDTs来自于早期残留或者施用农药后的长期风化残留。沉积物风险评估表明,巢湖东半湖表层沉积物中的有机氯农药存在一定的生态风险。     

Analysis of agricultural residues on tea using d-SPE sample preparation with GC-NCI-MS and UHPLC-MS/MS

This study presents new sample preparation and analytical procedures for the quantification of pesticides on processed tea leaves. The new method includes tea extraction and dispersive solid phase extraction (d-SPE) to prepare gas chromatography (GC) and ultrahigh-performance liquid chromatography (UHPLC)-ready samples, providing a fast and cost-effective solution for time-sensitive industrial analysis to fulfill regulatory requirements. Both GC-negative chemical ionization mass spectrometry (GC-NCI-MS) and UHPLC-tandem mass spectrometry (UHPLC-MS/MS) were employed to produce highly sensitive and reproducible data. Excellent limits of detection (typically below 1 μg/kg for GC and 10 μg/kg for UHPLC), wide linearity ranges, and good recoveries (mostly >70%) were achieved on the selected pesticides. Twenty-seven tea samples purchased from local grocery stores were analyzed using the newly developed methods. Among the pesticides analyzed, endosulfan sulfate and kelthane were the most frequently detected by GC-NCI-MS and imidacloprid and acetamiprid by UHPLC-MS/MS in these teas. The samples were found to be relatively clean, with <1 mg/kg of total pesticide residues. The organic-labeled teas were significantly cleaner than nonorganic ones. The cost per gram of tea did not correlate with pesticide residue levels detected.     

Occurrence,Degradation and Fate of Pesticides During Composting

This paper reviews the findings of research reported in the currently available literature regarding the occurrence and transformations of pesticides through the composting process and the use of compost. Part I summarizes the composting process, pesticides and mechanisms of pesticide degradation. Part II reviews research studies concerning the occurrence and fate of pesticides during composting. Investigations of pesticide residues in composting feedstocks and finished compost detected few of the target pesticides. The compounds that were found occurred at low concentrations. The majority of the compounds detected were insecticides in the organochlorine category, including chemicals that have been banned from use in the U.S. for many years. Generally, organophosphate and carbamate insecticides and most herbicides were rarely detected. Comparisons of pesticide concentrations before and after composting also showed organochlorine compounds to be most resistant to biodegradation during composting. With some exceptions, pesticides in other categories decomposed moderately well to very well. Studies that followed the mechanisms of degradation indicate that mineralization accounts for only a small portion of pesticide disappearance. Other prominent fates include partial degradation to secondary compounds, adsorption, humification, and volatilization. In general the research results suggest that the pattern of pesticide degradation during composting is similar to the degradation observed in soils. With a few important distinctions, composting can be considered a biologically active soil environment in which degradation is accelerated. However, as some studies noted, composting does not always speed the degradation of all pesticides. The nature of the pesticide, specific composting conditions and procedures, the microbial communities present, and the duration of composting affect the extent and the mechanisms of degradation.     

Literature Review: Occurrence,Degradation and Fate of Pesticides During Composting: Part II: Occurrence and Fate of Pesticides in Compost and Composting Systems

This paper reviews the findings of research reported in the currently available literature regarding the occurrence and transformations of pesticides through the composting process and the use of compost. Part I summarizes the composting process, pesticides and mechanisms of pesticide degradation. Part II reviews research studies concerning the occurrence and fate of pesticides during composting. Investigations of pesticide residues in composting feedstocks and finished compost detected few of the target pesticides. The compounds that were found occurred at low concentrations. The majority of the compounds detected were insecticides in the organochlorine category, including chemicals that have been banned from use in the U.S. for many years. Generally, organophosphate and carbamate insecticides and most herbicides were rarely detected. Comparisons of pesticide concentrations before and after composting also showed organochlorine compounds to be most resistant to biodegradation during composting. With some exceptions, pesticides in other categories decomposed moderately well to very well. Studies that followed the mechanisms of degradation indicate that mineralization accounts for only a small portion of pesticide disappearance. Other prominent fates include partial degradation to secondary compounds, adsorption, humification, and volatilization. In general the research results suggest that the pattern of pesticide degradation during composting is similar to the degradatiion observed in soils. With a few important distinctions, composting can be considered a biologically active soil environment in which degradation is accelerated. However, as some studies noted, composting does not always speed the degradation of all pesticides. The nature of the pesticide, specific composting conditions and procedures, the microbial communities present, and the duration of composting affect the extent and the mechanisms of degradation.     

Automated sample cleanup for pesticide multiresidue analysis. Part II--Design and evaluation of column chromatography module

An automated, continuous flow system is described for Florisil column chromatography of pesticide residues from food extracts. Evaluation of the system using 5 common organochlorine and organophosphorus pesticides in 2 crop matrices demonstrates essentially no difference in recovery or precision between automated and currently used manual analyses. The automated procedure uses only 20% of the solvents and adsorbents used in the manual procedure and is 3 times faster.     

新疆典型农业地区土壤中有机氯农药（OCPs）分布特征及风险评价

为加强有机氯农药(organic chlorinated pesticides,OCPs)的污染预防与控制,该研究分析了新疆典型农业地区有机氯农药的污染状况并对其进行了风险评价。采集表层土壤样品36个,分析其中15种OCPs的残留状况。六六六(Hexachlorocyclohexanes,HCHs)、滴滴涕(dichlorodiphenyltrichloroethanes,DDTs)、氯丹类化合物、硫丹和硫丹盐的质量分数范围分别是0.37～22.82、0.91～858.47、0.15～47.08ng/g、N.D(未检出)～16.27和N.D～73.83ng/g。对OCPs的来源进行分析,发现HCHs来源于历史上工业HCHs的使用或近期林丹的输入,DDT来源于工业品的违法使用和三氯杀螨醇的使用,研究区域氯丹存在新的来源,而硫丹新来源较少。通过主成分分析,从15种OCPs中提取了5个主成分,总方差解释量达到了79.93%。5个主成分分别归因于DDT和工业HCH s的使用、OCPs原料的差异以及研究区域内病虫害的发病特征等。利用灰色关联分析研究区域内OCPs污染状况,结果表明石河子污染水平最高,各研究区域土壤均需要采取进一步的污染控制措施。     

Simplified cleanup and gas chromatographic determination of organophosphorus pesticides in crops

A simple and efficient cleanup method for gas chromatographic determination of 23 organophosphorus pesticides in crops including onion is described. The sample was extracted with acetone. The extract was purified with coagulating solution, which contained ammonium chloride and phosphoric acid, and then filtered by suction. The filtrate was diluted with NaCl solution and reextracted with benzene. The organic layer was evaporated and injected into a gas chromatograph equipped with a flame photometric detector (FPD) and fused silica capillary columns (0.53 mm id) coated with silicone equivalent to OV-1701, OV-1, and SE-52. Onion extract, which contained FPD interferences, was cleaned up on a disposable silica cartridge. Recoveries of most organophosphorus pesticides from spiked crops: mandarin orange, tomato, spinach, sweet pepper, broccoli, lettuce, and onion at levels of 0.02-0.28 ppm, exceeded 80%, but the water-soluble pesticides dichlorvos and dimethoate gave poor recoveries in all crops; the nonpolar pesticides disulfoton, chlorpyrifos, fenthion, prothiophos, and leptophos were not recovered quantitatively in spinach, sweet pepper, broccoli, and lettuce. IBP, edifenphos, phosmet, and pyridaphenthion were not recovered from onion because of adsorption to the silica cartridge. The detection limits ranged from 1.25 to 17.5 ppb on a crop basis.     

Biodegradation kinetics of endosulfan by Fusarium ventricosum and a Pandoraea species

Endosulfan, classified as an organochlorine pesticide, is rated by the U.S. EPA as a Category 1 pesticide with extremely high acute toxicity. This study describes the biodegradation kinetics of endosulfan and the metabolic pathway utilized by Fusarium ventricosum and a Pandoraea sp. Complete disappearance of both alpha- and beta-endosulfan was observed during 12 days of incubation with F. ventricosum in flasks containing 100 mg L(-)(1) of endosulfan. The rate constants (k) for biodegradation of alpha- and beta-endosulfan by F. ventricosum using zero-order kinetics were 14.22 and 6.60 mg L(-)(1) day(-)(1), respectively. The Pandoraea sp. degraded about 95 and 100% of alpha- and beta-endosulfan, respectively, in 18 days of incubation in flasks spiked with 100 mg L(-)(1) of endosulfan. The rate constants (k) for biodegradation of alpha- and beta-endosulfan by the Pandoraea sp. were 8.19 and 3.78 mg L(-)(1) day(-)(1), respectively. Both fungal and bacterial strains formed less toxic endosulfan diol and endosulfan ether as metabolites during metabolism of endosulfan. The results of this study suggest that these novel strains may be used for the bioremediation of endosulfan-contaminated sites.     

设施大棚农药污染残留调查分析

利用弗罗里矽柱净化前处理方法,采用气相色谱（GC）方式,依据保留时间和特征离子丰度比,对辽宁某地区大棚内种植黄瓜及其叶片部分样品中的有机磷类农药（粉锈宁、乐果、敌敌畏、喹硫磷、辛硫磷、噻嗪酮、甲拌磷、马拉硫磷和对硫磷）,拟除虫菊酯类农药（百菌清、氯氰菊酯、氰戊菊酯、溴氰菊酯）的残留量进行检测分析。结果表明,有机磷类农药检出种类不固定,拟除虫菊酯类农药检出情况较为稳定,有机磷和拟除虫菊酯类农药在叶片中的检出率均略高于在果实中的检出率,且在不同时期不同大棚检出情况略有差异。针对当前农药施用现状,应多注意有机磷类和拟除虫菊酯类农药的使用和监管,对于有机磷类高毒性农药应减少或停止施用,中低毒性的拟除虫菊酯类农药应加强其作用的宣传并控制其使用量,避免追求高产量而盲目过量的施用,造成不必要的污染。     

Multiclass pesticide determination in olives and their processing factors in olive oil: comparison of different olive oil extraction systems

The processing factors (pesticide concentration found in olive oil/pesticide concentration found in olives) of azinphos methyl, chlorpyrifos, lambda-cyhalothrin, deltamethrin, diazinon, dimethoate, endosulfan, and fenthion were determined in olive oil production process in various laboratory-scale olive oil extractions based on three- or two-phase centrifugation systems in comparison with samples collected during olive oil extractions in conventional olive mills located at different olive oil production areas in Greece. Pesticide analyses were performed using a multiresidue method developed in our laboratory for the determination of different insecticides and herbicides in olive oil by solid-phase extraction techniques coupled to gas chromatography detection (electron capture detection and nitrogen phosphorus detection), optimized, and validated for olive fruits sample preparation. Processing factors were found to vary among the different pesticides studied. Water addition in the oil extraction procedure (as in a three-phase centrifugation system) was found to decrease the processing factors of dimethoate, alpha-endosulfan, diazinon, and chlorpyrifos, whereas those of fenthion, azinphos methyl, beta-endosulfan, lambda-cyhalothrin, and deltamethrin residues were not affected. The water content of olives processed was found to proportionally affect pesticide processing factors. Fenthion sulfoxide and endosulfan sulfate were the major metabolites of fenthion and endosulfan, respectively, that were detected in laboratory-produced olive oils, but only the concentration of fenthion sulfoxide was found to increase with the increase of water addition in the olive oil extraction process.     

Comparison of Two Procedures for Extraction and Clean-up ofOrganophosphorus and Pyrethroid Pesticides in Sediment

Two procedures were compared for extraction and clean-up of 20 organophosphorus and 19 pyrethroid pesticidesin sediment to identify the more effective procedure for groups of pesticides or individual compounds. In Procedure I,methanol/water and n-hexane were used for extraction, and 1:10 (v/v) dichloromethane in n-hexane and acetone wereused as eluents for eluting the analyte through the cartridge, with one evaporating steps on a rotary evaporator and twoeluting steps on the cartridge, n-hexane/acetone (2:1, v/v) was used for extraction and elution in Procedure II with oneevaporating step on a rotary evaporator and one eluting step oll the cartridge. All extractions were performed underan ultrasonic bath and gas chromatography and mass spectrometry were utilized for measurements. Procedure II wasdeveloped as a rapid, timesaving, less costly and safer substitute for Procedure I which was an old method. ProcedureII was more effective for almost all the organophosphorus pesticides tested and 11 of the 19 pyrethroid pesticides, whileProcedure I was more appropriate for analysis of 5 pyrethroid pesticides. However, recoveries of most pyrethroid pesticideswere fairly low. Thus, further studies should focus on adjustment and formulation of solvents for more efficient extractionand clean-up of pyrethroid pesticides from sediment samples.     

The Luke et al. method for determining multipesticide residues in fruits and vegetables: collaborative study

Ten laboratories analyzed unfortified and fortified samples of lettuce, tomatoes, and strawberries for organochlorine and organophosphorus pesticides by applicable portions of the comprehensive multipesticide method of Luke et al. The 3 crops were fortified with 6 pesticides, alpha-BHC, dieldrin, chlorpyrifos, acephate, omethoate, and monocrotophos, each at 3 levels per crop. Included in the 54 fortifications were 16 pairs of blind duplicates: same pesticide, crop, and level. Recoveries were calculated by area comparisons with known reference materials, using the responses obtained from 2 separate element-specific gas chromatographic (GC) systems. The organochlorine pesticides were chromatographed on a methyl silicone column and detected with a Hall 700A electrolytic conductivity detector, and the organophosphorus pesticides were determined with a flame photometric detector after being chromatographed on a specified DEGS column material. Chlorpyrifos was quantitated on both GC systems. Mean recoveries ranged from 82.6% for acephate fortified at 0.5000 ppm in strawberries to 118.1% for 0.0636 ppm fortification of chlorpyrifos in lettuce. Interlaboratory coefficients of variation ranged from 4.0% for 0.6360 ppm fortification of chlorpyrifos in tomatoes to 17.8% for the 0.0636 ppm chlorpyrifos level in lettuce. The procedure features essentially no cleanup before GC and proved comparable to existing multiresidue methods for pesticides of the class types studied, as evidenced by the intra- and interlaboratory measurements of precision and recoveries obtained. The method with the 2 GC systems has been adopted official first action.     

Collaborative study of the determination of endosulfan, endosulfan sulfate, tetrasul, and tetradifon residues in fresh fruits and vegetables.

A method for the determination of endosulfan I, endosulfan II, endosulfan sulfate, tetrasul, and tetradifon residues in fresh fruits and vegetables was studied collaboratively. The method consists of extraction of the product with acetonitrile, ether and elution from a Florisil column with mixtures of hexane, methylene chloride, and acetonitrile, and determination of the residues by electron capture gas-liquid chromatography. This method was studied by 8 collaborators, using apples and cucumbers as the samples. Each sample was spiked with 2 levels of each pesticide. The average per cent recoveries for apples and cucumbers, respectively were as follows: endosulfan I, 103.6 and 101.5; endosulfan II, 102.9 and 100.0; endosulfan sulfate, 100.9 and 92.9; tetrasul, 98.8 and 102.2; and tetradifon, 106.4 and 101.9. The method has been adopted as official first action.     

An evaluation of methods for measurement of pesticides in sorption experiments

Abstract

The concentration of four pesticides (2,4‐D, atrazine, phorate, and terbufos) in soil solution during sorption experiments was measured using UV spectrophotometry, Gas Liquid Chromatography (GLC), High Performance Liquid Chromatography (HPLC), and radiotracer technique. The presence of water soluble organic matter in soil solution interfered with the measurement of pesticide using the UV spectrophotometry. The use of GLC, HPLC, and radiotracer technique involving ¹⁴C gave a good estimate of the concentration of pesticide in soil solution. The pesticide remaining in soil can be quantitatively analyzed by extracting with a scintillation solution containing an organic solvent such as toluene or dioxane. Among the various centrifuge tubes glass tubes with Teflon caps sorbed negligible amount of pesticides and these tubes can be used for the sorption measurements.